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Pwo DNA Polymerase

erial and if after PCR single DNA molecules are analyzed, PCR artifacts can be a significant problem. Fidelity of DNA polymerization is for instance important for:
  • cloning of PCR products
  • study of allelic polymorphism in individual RNA transcripts (1, 2)
  • characterisation of the allelic stage of single cells (3) or single DNA molecules (4, 5)
  • characterisation of rare mutations in tissue (6)
  • characterisation of a population of cells in culture
Operating parameters
  • Thermal stability: Pwo DNA Polymerase is stable for >2 h at 95C
  • Divalent ion requirement: Mg2+ (standard concentration, 2.00 mM)
  • dNTP concentration: at least 200 M for each dNTP
    Caution: Lower deoxynucleotide concentrations can increase fidelity but can also activate the 3'5' exonuclease proofreading activity, which might degrade prime
Experimental results Key advantages
  • Maximum PCR fidelity, because the proofreading activity of Pwo DNA Polymerase excises misincorporated nucleotides
  • High PCR yield, because the polymerase is very resistant to the high denaturing temperatures used in PCR
  • Simplifies cloning of PCR products because it generates blunt-ended products which can be directly used f
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Recommend to use at 1000X dilutions for cell culture (e.g. Add 1 ul to 1 ml of culture medium.)...
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